Generation and neutralization of pseudovirions of human papillomavirus type 33
| Autor: | Martin Sapp, F Unckell, Rolf E. Streeck |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
viruses
Immunology Biology Antibodies Viral complex mixtures Microbiology Neutralization law.invention chemistry.chemical_compound Capsid Plasmid Neutralization Tests law Virology Animals Deoxyribonuclease I Humans Antigens Viral Papillomaviridae Antiserum Virus Assembly Virion virus diseases Oncogene Proteins Viral Molecular biology In vitro Titer chemistry Insect Science COS Cells DNA Viral Recombinant DNA Capsid Proteins DNA Research Article |
| Zdroj: | Journal of Virology. 71:2934-2939 |
| ISSN: | 1098-5514 0022-538X |
| DOI: | 10.1128/jvi.71.4.2934-2939.1997 |
| Popis: | Since human papillomaviruses (HPV) cannot be propagated in cell culture, the generation of infectious virions in vitro is a highly desirable goal. Here we report that pseudovirions can be generated by the assembly of virus-like particles (VLPs) in COS-7 cells containing multiple copies of a marker plasmid. Using recombinant vaccinia viruses, we have obtained spherical VLPs of HPV type 33 (HPV-33) which fractionate into heavy and light VLPs in cesium chloride density gradients. VLPs in the heavy fraction (1.31 g/cm3) carry the plasmid in DNase-resistant form and are capable of transferring the genetic marker located on the plasmid to COS-7 cells in a DNase-resistant way (pseudoinfection). The minor capsid protein L2 is not required for encapsidation but is essential for efficient pseudoinfection. Antiserum to HPV-33 VLPs inhibits VLP-mediated DNA transfer with high efficiency. Antisera to VLPs of HPV-18 and HPV-16 are not neutralizing, although the HPV-16 antiserum exhibited some cross-reactivity with HPV-33 VLPs in an enzyme-linked immunosorbent assay. In a cell binding assay, the titer of the HPV-33 VLP antiserum was 1:200 compared to the neutralization titer of 1:10(5). This indicates that neutralization is essentially due to the inhibition of cellular processes after VLP binding to cells. The encapsidation of marker plasmids into VLPs provides a sensitive and fast assay for the evaluation of neutralizing potentials of antisera against papillomavirus infections. |
| Databáze: | OpenAIRE |
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